Bagriaçik E U, Armstrong M D, Okabe M, Klein J R
Department of Biological Science and the Mervin Bovaird Center for Studies in Molecular Biology and Biotechnology, University of Tulsa, Tulsa, OK 74104, USA.
Int Immunol. 1999 Oct;11(10):1651-62. doi: 10.1093/intimm/11.10.1651.
Three mAb (R2/60, S7 and 1B11) were used to study the expression of murine CD43 on peripheral T cells and intestinal intraepithelial lymphocytes (IEL) from normal mice, and from mice during acute graft versus host disease (GVHD). In the spleen, essentially all T cells expressed the R2/60 and S7 antigens, whereas the 1B11 antigen was expressed on about half of the CD8(+) cells and approximately 15% of CD4(+) T cells. Interestingly, a significant proportion of resting splenic B cells expressed the 1B11 and R2/60 antigens, but not the S7 antigen. The majority of IEL expressed R2/60 antigen; however, the S7 and 1B11 markers were differentially expressed on CD8alpha, CD8beta, TCRalphabeta and TCRgammadelta cells. Immunoprecipitation and Western blotting analyses identified characteristic 115 and 130 kDa reactive components from IEL lysates with mAb S7 and 1B11 respectively, and reactivity to both molecular entities by mAb R2/60. During acute intestinal GVHD induced by injecting CB6F(1) athymic nude mice with spleen cells from C57BL/6 enhanced-green fluorescent protein transgenic mice, 80-90% of donor T cells in the intestine epithelium expressed all CD43 isoforms; however, the level of expression of the 130 kDa CD43 antigen increased significantly and the level of the 115 kDa antigen decreased on GVHD donor T cells compared to cells at the time of transfer. Using EL4 cells, a similar shift in the expression of CD43 isoforms occurred experimentally following treatment with neuraminidase, suggesting that the type of CD43 isoform expressed on T cells is strongly influenced by conditions which affect membrane charge. The significance of these findings for intestinal immunopathology is discussed.
使用三种单克隆抗体(R2/60、S7和1B11)研究正常小鼠以及急性移植物抗宿主病(GVHD)小鼠外周血T细胞和肠道上皮内淋巴细胞(IEL)上小鼠CD43的表达情况。在脾脏中,基本上所有T细胞都表达R2/60和S7抗原,而1B11抗原在约一半的CD8(+)细胞和约15%的CD4(+) T细胞上表达。有趣的是,相当一部分静息脾B细胞表达1B11和R2/60抗原,但不表达S7抗原。大多数IEL表达R2/60抗原;然而,S7和1B11标志物在CD8α、CD8β、TCRαβ和TCRγδ细胞上的表达存在差异。免疫沉淀和蛋白质印迹分析分别从IEL裂解物中鉴定出与单克隆抗体S7和1B11反应的特征性115 kDa和130 kDa反应成分,以及与单克隆抗体R2/60对这两种分子实体的反应性。在用C57BL/6增强型绿色荧光蛋白转基因小鼠的脾细胞注射CB6F(1)无胸腺裸鼠诱导的急性肠道GVHD期间,肠道上皮中80 - 90%的供体T细胞表达所有CD43异构体;然而,与转移时的细胞相比,GVHD供体T细胞上130 kDa CD43抗原的表达水平显著增加,而115 kDa抗原的水平降低。使用EL4细胞,在用神经氨酸酶处理后实验上出现了类似的CD43异构体表达变化,表明影响膜电荷的条件对T细胞上表达的CD43异构体类型有强烈影响。讨论了这些发现对肠道免疫病理学的意义。